LOCUS       sequence-233991-        5536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allelic exchange plasmid with PheS* counter-selection for 
            Staphylococcus aureus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5536)
  AUTHORS   Schuster CF, Howard SA, Grundling A
  TITLE     Use of the counter selectable marker PheS* for genome engineering in
            Staphylococcus aureus.
  JOURNAL   Microbiology. 2019 Apr 3. doi: 10.1099/mic.0.000791.
  PUBMED    30942689
REFERENCE   2  (bases 1 to 5536)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microbiology. 2019 Apr 3. doi: 10.1099/mic.0.000791."
COMMENT     Sequence Label: pIMAY*
FEATURES             Location/Qualifiers
     source          1..5536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2388..2399
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     oriT            3949..4058
                     /note="incP origin of transfer"
     rep_origin      4200..4745
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4646..4665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4862..4879
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5252..5271
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5252..5270
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    5279..5386
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     5303..5319
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5303..5319
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5353..5369)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5354..5370)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(5399..5417)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 atattataat tatcaaagag aggggtggca aacggtattt ggcattatta ggttaaaaaa
       61 tgtagaagga gagtgaaacc catgaacttt aataaaattg atttagacaa ttggaagaga
      121 aaagagatat ttaatcatta tttgaaccaa caaacgactt ttagtataac cacagaaatt
      181 gatattagtg ttttataccg aaacataaaa caagaaggat ataaatttta ccctgcattt
      241 attttcttag tgacaagggt gataaactca aatacagctt ttagaactgg ttacaatagc
      301 gacggagagt taggttattg ggataagtta gagccacttt atacaatttt tgatggtgta
      361 tctaaaacat tctctggtat ttggactcct gtaaagaatg acttcaaaga gttttatgat
      421 ttataccttt ctgatgtaga gaaatataat ggttcgggga aattgtttcc caaaacacct
      481 atacctgaaa atgctttttc tctttctatt attccatgga cttcatttac tgggtttaac
      541 ttaaatatca ataataatag taattacctt ctacccatta ttacagcagg aaaattcatt
      601 aataaaggta attcaatata tttaccgcta tctttacagg tacatcattc tgtttgtgat
      661 ggttatcatg caggattgtt tatgaactct attcaggaat tgtcagatag gcctaatgac
      721 tggcttttat aatatgagat aatgccgact gtacttttta cagtcggttt tctaatgtca
      781 ctaacctgcc ccgttagttg acatatgatc atcataattc tgtctcatta tataacatcc
      841 tccatacctt ctattataga atgtattgct attaatcgca acatcaaacc aaaataaaaa
      901 cccccttcga ctttcgtcag ggggctttta tttattcaat aatccctcct ctcaataaat
      961 ctattgttgt acttaattca acttccattt ctctgtatct ttcaatacgc tcttttaagt
     1021 ccttaatttc tttttttaat tcctcatttt cagcaaataa ctctttttct ttgtttgtca
     1081 ttttatttcc cccgtttcag catcaagaac ctttgcataa cttgctctat atccacactg
     1141 ataattgccc tcaaaccata atctaaaggc gctagagttt gttgaaacaa tatcttttac
     1201 atcattcgta tttaaaattc caaactccgc tcccctaagg cgaataaaag ccattaaatc
     1261 ttttgtattt accaaattat agtcatccac tatatctaaa agtaaattct tcaattctct
     1321 tttttggctt tcatcaagtg ttatatagcg gtcaatatca aaatcattaa tgttcaaaat
     1381 atcttttttg tcgtatatat gtttattctt agcaatagcg tcctttgatt catgagtcaa
     1441 atattcatat gaacctttga tataatcaag tatctcaaca tgagcaactg aactattccc
     1501 caattttcgc ttaatcttgt tcctaacgct ttctattgtt acaggatttc gtgcaatata
     1561 tataacgtga tagtgtggtt ttttatagtg ctttccattt tgtataatat tactattatt
     1621 ccatgtatct ttatcttttt tttcgtccat atcgtgtaaa ggactgacag ccatagatac
     1681 gcccaaactc tctaattttt ctttccaatc attaggaatt gagtcaggat ataataaaaa
     1741 tccaaaattt ctagctttag tatttttaat agccatgata taattacctt atcaaaaaca
     1801 agtagcgaaa actcgtatcc ttctaaaaac gcgagctttc gcttattttt tttgttctga
     1861 ttcctttctt gcatattctt ctatagctaa cgccgcaacc gcagattttg aaaaaccttt
     1921 ttgtttcgcc atatctgtta attttttatc ttgctctttt gtcagagaaa tcataactct
     1981 ttttttcgat tctgaaatca ccatttaaaa aactccaatc aaataatttt ataaagttag
     2041 tgtatcactt tgtaatcata aaaacaacaa taaagctact taaatataga tttataaaaa
     2101 acgttggcga aaacgttggc gattcgttgg cgattgaaaa accccttaaa cccttgagcc
     2161 agttgggata gagcgttttt ggcacaaaaa ttggcactcg gcacttaatg gggggtcgta
     2221 gtacggaagc aaaattcgct tcctttcccc ccattttttt ccaaattcca aatttttttc
     2281 aaaaattttc cagcgctacc gctcggcaaa attgcaagca atttttaaaa tcaaacccat
     2341 gagggaattt cattccctca tactcccttg agcctcctcc aaccgaaata gaagggcgct
     2401 gcgcttatta tttcattcag tcatcggctt tcataatcta acagacaaca tcttcgctgc
     2461 aaagccacgc tacgctcaag ggcttttacg ctacgataac gcctgtttta acgattatgc
     2521 cgataactaa acgaaataaa cgctaaaacg catgcggatc ccgagttcat gaaaaactaa
     2581 aaaaaatatt gacactctat cattgataga gcttaattaa aataagcttg atatcgaatt
     2641 aaggaggaaa aaaaatgtca gaacaacaaa caatgtctga attaaaacag caagcattag
     2701 tagatataaa cgaagcaaat gacgaaagag ctttacaaga agtgaaagtc aaatatttgg
     2761 gaaagaaagg ttctgttagt ggattaatga aattgatgaa agacttacca aatgaagata
     2821 aaccagcttt tggtcagaaa gttaatgaac taagacaaac tatacagaat gaacttgatg
     2881 agagacaaca aatgctagtg aaagaaaaat taaacaaaca gctagcagaa gaaactattg
     2941 atgtctcatt accaggaaga catattgaaa tcggatctaa acatccttta acgcgaacga
     3001 ttgaagaaat cgaagactta tttttaggtc ttggttatga aatagtaaac ggatatgagg
     3061 ttgaacaaga tcattacaac ttcgagatgt taaatcttcc aaaatctcat ccagcacgag
     3121 atatgcaaga tagtttctac ataacagatg aaattttatt acgaacgcat acttctccag
     3181 tacaagctag aactatggaa tcaagacatg gacaaggtcc tgtaaaaatt atttgtccag
     3241 gtaaagttta cagaagagat tctgatgatg cgactcattc acatcaattt acacaaatag
     3301 aaggattagt tgttgataaa aatgtgaaga tgagtgatct taaaggaaca ttggagttac
     3361 ttgcaaaaaa actatttgga gctgatcgag aaataagatt aagacctagt tattttccat
     3421 ttacagaacc atctgtagaa gtagatgtat cttgttttaa atgtaaaggc aaaggttgca
     3481 atgtttgtaa acacacggga tggattgaaa tcttaggtgc aggcatggtt cacccaaatg
     3541 tcttagagat ggctggtttt gactcaagtg agtatagtgg atttggtttt ggcatgggtc
     3601 cggatagaat tgctatgtta aaatacggaa tcgaagatat tcgacacttc tacactaatg
     3661 atgtacgttt tttagatcaa tttaaagcag ttgaagatcg tggagatatg taaggtacca
     3721 gatctcctct cgcctgtccc ctcagttcag taatttcctg catttgcctg tttccagtcg
     3781 gtagatattc cacaaaacag cagggaagca gcgcttttcc gctgcataac cctgcttcgg
     3841 ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg
     3901 ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg
     3961 tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc
     4021 ggtgctcaac gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga
     4081 gggcaagcgg cggagaatta caacttatat cgtatggggc tgacttcagg tgctacattt
     4141 gaagagataa attgcactga aatctagaaa tattttatct gattaataag atgatcttct
     4201 tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag
     4261 ggcggttttt cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga
     4321 ggagcgcagt caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag
     4381 actaactcct ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc
     4441 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg
     4501 ttcgtgcata cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg
     4561 aatgagacaa acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag
     4621 gagagcgcac gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4681 ttcgccacca ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat
     4741 ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc
     4801 caggaaatct ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg
     4861 tagcgagtca gtgagcgagg aagcggaata tatcctgtat cacatattct gctgacgcac
     4921 cggtgcagcc ttttttctcc tgccacatga agcacttcac tgacaccctc atcagtgcca
     4981 acatagtaag ccagtataca ctccgctagc gctgatgtcc ggcggtgctt ttgccgttac
     5041 gcaccacccc gtcagtagct gaacaggagg gacagctgat agaaacagaa gccactggag
     5101 cacctcaaaa acaccatcat acactaaatc agtaagttgg cagcatcacc cgacgcactt
     5161 tgcgccgaat aaatacctgt gacggaagat cacttcgcag aataaataaa tcctggtgtc
     5221 cctgttgata ccgggaagcc ctgggacgtc gtaatacgac tcactatagg gcgaattgga
     5281 gctccaccgc ggtggcggcc gctctagaac tagtggatcc cccgggctgc aggaattcga
     5341 tatcaagctt atcgataccg tcgacctcga gggggggccc ggtacccagc ttttgttccc
     5401 tttagtgagg gttaattatc ccattatgct ttggcagttt attcttgaca tgtagtgagg
     5461 gggctggtat aatcacatac ggccgataaa gcaagcatat aatattgcgt ttcatcttta
     5521 gaagcgaatt tcgcca
//
